Genade: Killi-data News (Winter) KILLI-DATA INTERNATIONAL Killi-Data News Volume 2, Issue 4, Pages 68–97, December 2017

ISSN 2495-330X Editorial Executive editor: Tyrone Genade (killifl[email protected]) Valued readers, it is with a heavy heart that I inform you that Editor: Jean H. Huber this is the last issue of Killi-Data News. The good news is that Contributors: Hristo Hristov (photo), Jean Huber (text), Bela Nagy (photos), we will be back as Killifish Research Review. Andy Patel (text), Bettina Reichenbacher (text), Andrew Thompson (text), Stefano Valdesalici (text), Frans Vermeulen (text and photos) The dissolution of Killi-Data International created a prob- lem: how can the newsletter of a defunct organization live Contents on without that organization? But other additional problems were building in the background. The first issue numbered Editorial ...... 68 15 pages. The previous issue was 28 pages. The number of Erratum ...... 69 killifish related papers is increasing while time on our end In the News ...... 69 (the editorial team) is running out. It takes a lot of time to Review of new research publications ...... 69 read the papers and put all the contributions together into New Book ...... 69 the newsletter. Were it not for the people writing us to inform Systematics, Taxonomy & Distribution ...... 70 us that they appreciate our work we would probably give up. Killifish Biology ...... 79 So we intend to continue in a new format. We are starting Posters & Conference Abstracts ...... 94 the journal afresh on Elsevier’s (formerly Bepress’s) Digital Interesting Research ...... 96 Commons platform which will enable us to publish reviews and summaries ad hoc, with open access and operating as a journal with a modicum of peer review. At this time we don’t yet have a link to the new journal page but this will be mailed out when we launch early next year. In the interim we hope you, the readers, will help us and come on board in a reviewer or contributor capacity. We need people with expertise in physiology, molecular phylogenetics, taxonomy, toxicology and ecology. As we are rebranding as a Killi-Data International retains copyright and distribution rights to all contents of Killi- killifish review journal we are after review articles on killifish- Data News unless otherwise claimed by the submitting author. related research. Submissions will be much appreciated.

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 68 1 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 There is a diverse assortment of papers reviewed in this Review of New Research Publications issue. The special edition of Developmental Dynamics on an- nual fish has provided a lot of material (some of it was given New Books in the previous issue). We draw your attention to Podrabsky and Arezo’s editorial which lays out the need for research Intermittent Rivers and Ephemeral Streams: Ecology and on annual fish. We also have two papers that are relevant to Management. Editors: Datry T, Bonada N and Boulton your health: the effect of glyphosate on Aphanius and arsenic A. Elsevier Science & Technology Books, 2017, ISBN: 978- on Fundulus. We also have new descriptions and molecular 0-12-803835-2. 622 pages. DOI https://doi.org/10.1016/ systematics work. We are fortunate to have Bettina Reichen- B978-0-12-803835-2.09997-6. bacher reviewing the recent description of the new killifish This book has several chapters of interest to killifish hob- genus, Pseudorestias. byists and researchers: We hope you enjoy this issue and catch up with us in the Chapter 4.10: Genetic, Evolutionary, and Biogeographical new year at our new home on the Digital Commons. Processes in Intermittent Rivers and Ephemeral Streams by [Tyrone Genade] Bonada N, Carlson, S, Datry, T, Finn, DS, Leigh, C and Lytle DA, Monaghan MT, Tedesco PA. Chapter 4.5: The Biota of Intermittent Rivers and Erratum Ephemeral Streams: Fishes by Kerezsy A, Gido K, Magalhães F and Skelton PH. On page 57 of the previous issue I refered to “Austrolebias lim- naeus enters Diapause II at 15 days post fertilization and. . . ‘’ Ichthyofauna of the Itimbiri, Aruwimi, and Lindi/Tshopo that should be Austrofundulus limnaeus. Rivers (Congo basin): diversity and distribution patterns. Decru E; Vreven E; Danadu C; Walanga A; Mambo T; & Snoeks J. Acta Ichthyologica et Piscatoria, 47:225–247, 2017. URL In The News https://www.aiep.pl/volumes/2010/8_3/txt/txt_02.php This is a catalog of ichthyofauna collected from the Itim- Killifish have made their way into the news in several articles: biri, Aruwimi, and Lindi/Tshopo Rivers. It lists many species, http://www.alraimedia.com/ar/article/local/2017/10/ including 19 species of killifish. 30/801302/nr/nc A story about Aphanius Kuwait’s Jahra [Tyrone Genade] Reserve and efforts to conserve it and its biodiversity.

http://www.news-gazette.com/news/business/2017-10-22/ wired-becky-fuller.html Becky Fuller’s research on Lucania goodei has made the news.

https://www.israel21c.org/tiny-fish-swims-to-israel-to- help-unlock-mystery-of-aging/ Itamar Harel has started a Nothobranchius research lab in Jerusalem. https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 69 2 Genade: Killi-data News (Winter) Systematics, Taxonomy & Distribution et al. (2016). A different phylogenetic interpretation was pub- lished by Costa (1997) based on a data matrix comprising 65 morphological characters. According to his analysis, Orestias Morphological and taxonomic descriptions of a new genus would be sister to a large clade comprising Aphanius + New and species of killifishes (Teleostei: Cyprinodontiformes) World Cyprinodontini, which is actually more in line with the from the high Andes of northern Chile. Arratia G; Vila I; Lam disjunct biogeography of Orestias and Aphanius and addition- N; Guerrero CJ; & Quezada-Romegialli C. PLOS ONE, 12:1–36, ally confirmed by recent molecular-based work (e.g. Reznick 2017. DOI https://doi.org/10.1371/journal.pone.0181989. et al. 2017). Arratia et al. (2017) provide now a new list of This is a major work dedicated to the description of a new characters diagnosing the Orestiini, containing Orestias and killifish genus and species from the southern Chilean Alti- Pseudorestias lirimensis as sole members, and thus contribute plano of the Andean mountains. Its name is Pseudorestias to a much improved understanding of this tribe. lirimensis, and it comes from the wetlands and streams of the As it is the case for many endemic killifish species, also Chancacolla River at about 4,000 m height (a.s.l.). In previous Pseudorestias lirimensis must be considered as critically en- work, Pseudorestias lirimensis had been tentatively assigned dangered. This threat is not only because of its small habitat to Orestias cf. agassii. However, numerous differences in the and reduced distribution, but also due to water demands and morphological characters, coloration and karyotype provide climate change in the region. Its recognition as a new genus compelling evidence that P. lirimensis is completely differ- and species will hopefully help to develop new conservation ent from all other species of Orestias. Its autapomorphies strategies for the southern Altiplano, from where, besides P. comprise unique configurations of several structures, i.e. (i) lirimensis, also six endemic species of Orestias are known. the Meckel cartilage, (ii) the basibranchials, (iii) the second [Bettina Reichenbacher, Department of Earth and Envi- pharyngobranchial, (iv) the ceratohyal, and (v) the dorsal as ronmental Sciences, Ludwig-Maximilians University, Munich, well as the (vi) anal radials. In addition, P. lirimensis displays Germany.] strong sexual dimorphism, with females being nearly twice as large and long than males, while males show absence of neu- Deep and concordant subdivisions in the self-fertilizing man- romast lines and reduced squamation on the head (amongst grove killifishes (Kryptolebias) revealed by nuclear and others). The authors provide detailed and thorough descrip- mtDNA markers. Tatarenkov A; Lima SMQ; Earley RL; tions of P. lirimensis, together with excellent drawings and Berbel-Filho WM; Vermeulen FBM; Taylor DS; Marson K; photos. They suggest that the emergence of the new genus and Turner BJ; & Avise JC. Biological Journal of the Linnean So- species may have resulted from Miocene-Pliocene vicariance ciety, 122:558–578, 2017. DOI http://dx.doi.org/10.1093/ events and the isolation of the Chancacolla River ecosystem biolinnean/blx103. since then. This is a high impact paper. It calls into question current Moreover, this work contributes to solve the debate understanding of the fishes we call Kryptolebias hermaphrodi- whether a sister-relationship exists between the Old-World tus (sensu Costa, 2011) and marmoratus and sets the stage killifish Aphanius and the New-World killifish Orestias. Based for multiple descriptions of new species in a cryptic species on morphological evidence, such a relationship was proposed complex and the reinstatement of old species long ago syn- by Parenti (1981, 1984), and has also been adopted in Nelson onymised with K. marmoratus.

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 70 3 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 The authors analyzed 734 specimens from 33 locations through to Venezuela, also most of the Caribbean islands from Florida, USA to Fazenda River, Brazil. K. caudomargina- haven’t been sampled making it difficult to delineate the bor- tus (sensu Costa) was not included in this study. This study ders between the species. The authors adopt a conservative restricted itself to the topic K. hermaphroditus and K. marmora- analysis of the data, interpreting it as support for K. marmora- tus. 33 nuclear microsatelite DNA sequences were studied in tus, K. hermaphroditus (sensu Costa, 2011) and K. bonairen- addition to regions of the mitochondrial cytochrome oxidase sis/heyei. The authors adopt the name bonairensis for the subunit I and cytochrome B genes. Microsatellite, cytochrome Central Clade because the island of Bonaire has been sam- oxidase subunit I and cytochrome B data revealed three major pled. The taxon heyei (from the Island of Saona, Dominican clades (Northern, Central and Southern). The Northern and Republic) is older than bonairensis. Should samples from the Central Clades are both derived from the Southern Clade, Island of Saona group with the Central Clade it would have which itself is hypothesized to have its origin in northern to be given the name heyei or the Antilles Clade be separated South America. Within the Southern Clade are three smaller out from the Bonaire-Aruba Clade as a distinct species. The groupings: two around the north east corner of Brazil and the authors discuss at great length the decision of whether to go third around Rio de Janeiro. Within the central clade there are with one, two or three species. They write “ The decision again three groups: the Antilles, Bonaire- Aruba and Panama. on taxonomic status of the three identified lineages of Kryp- In the Northern Clade the Honduras-Belize clade separates tolebias is thus somewhat subjective and largely determined out from the Florida-Bahamas clade. These sub-clades have by the adherence of a researcher to one or another school lower support than the three major clades. of thought, BSC or PSC. Anecdotally, even within the small group of co-authors of this study, we could not come to con- sensus on this subject.” The authors are adamant on the data for the Southern Clade supporting a one species hypothesis of K. hermaphroditus only. The authors conclude that “Our find- ing of two highly distinct Caribbean lineages—Northern and Central—raises intriguing questions. Are there significant (but hitherto unrecognized) biological differences between the lineages? Are they (or were they ever) in direct contact? Does introgression or displacement occur in putative contact zones? The reclusive K. marmoratus is still far from revealing all of its secrets.” Indeed, this paper is probably the beginning for systematic Kryptolebias research. As an aside, Guy Tiriault of the Island of Guadeloupe, was generous enough to supply us with samples of Kryptolebias Photo of male K. marmoratus Dangriga and hermpahrodite K. from the island of Guadeloupe. These have been entered bonairensis by Frans Vermeulen© into the DNA bank and samples have already been sent to Dr Andrei Tatarenkov and analyzed, identifying them with the There is a dearth of specimens from northern Brazil Central Clade. Live fish of this collection are being distributed https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 71 4 Genade: Killi-data News (Winter) as Kryptolebias sp. cf. bonairensis Bon Gout GT-15. I had set- tled on this designation based on the clear genetic separation of these fish from K. marmoratus and K. hermaphroditus. I am not committed to the bonairensis name for these, and suspect that in the end heyei will win out for the Antilles clade. I leave this matter to the professional taxonomists to settle. Also, fish from Puerto Rico are also of the Central clade and are not marmoratus. If you collect Kryptolebias while on holiday in South/Central America or the Caribbean, please preserve a specimen for DNA analysis! [Tyrone Genade]

Aphanius kruppi, a new killifish from Oman with com- ments on the A. dispar species group (Cyprinodontiformes: Aphaniidae) . Freyhof J; Weissenbacher A; & Geiger M. Zootaxa, 4338:557–573, 2017. DOI http://dx.doi.org/10. 11646/zootaxa.4338.3.10. This is a very important paper in the field of Aphanius re- search and should be read by hobbyists that keep Aphanius as well. This paper presents a description for Aphanius kruppi as well as provides an identification key for the Aphanius dispar species group. It also presents a DNA phylogenetic scheme for the species group which raises the very important question of just what is A. dispar? A. kruppi is named for Faheed Krupp for his many valu- able contributions to the explorations of the freshwater fishes of the Middle East. The specimens used to describe the species were obtained on a hobbyist collection trip to Oman in 2015 that were raised by the Zoo of Vienna. The fish comes from the Al Mudayrib spring and Wadi Bani Khalid at Sayh al Hayl Photos of male and female Aphanius kruppi from Al Mudayrib (both sites are in Wadi al Batha which drains into the Arabian (top) and Wadi Bani Khalid (bottom), Oman. Photo taken from Sea north of Al Ashkharah). A. kruppi has a stable and distinct Freyhof et al©. phenotype that allows it to be easily discerned from other species. It differs from sirhani in females haveing 7–12 short blotches as well as three black bars on the caudal fin. It differs bars on its flank and males lacking a black dorsal-fin margin from A. dispar, richardsoni and stoliczkanus by having black of and 5–10 irregularly set and shaped bands of spots or dark gray bars on the flank and a diamond-shaped

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 72 5 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 having 9–14 brown or grey bars on the flank. It differs from A. furcatus by having a truncate caudal fin and the body that is completely covered with scales (A. furcatus has a partially scaleless body). A. stiassnyae does not have the upturned lower jaw. The authors consider A. kruppi as a threatened species due to its small distribution and very small populations.

Photo of male and female Aphanius cf. dispar from Atlit, Israel. Photo taken from Freyhof et al©. The molecular analysis of the cytochrome oxidase I gene produced a phylogeny with strong support but which did not Photos of male and female Aphanius cf. kruppi from Al-Hoota, align with the current conception of the species. A. dispar Oman (top) and Aphanius stoliczkanus fromWadi Fanja, Oman breaks into three distinct clades (Egypt-Israel, Eritrea and (bottom). Photo taken from Freyhof et al©. Saudi Arabia). A. kruppi and A. stoliczkanus seem to have hybridized at Khor Kalba nature reserve (United Arab Emi- or vertically-elongate black or brown blotch at the base of rates), Fanja and Al Hoota, Oman (which shows an interme- the caudal-fin of females. The dorsal and anal fins are short diate color pattern between the species), but in general A. relative to these three species. It differs from A. ginaonis by stoliczkanus has a discrete and wide range. It is unlikely that https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 73 6 Genade: Killi-data News (Winter) A. dispar exists in Iran, instead, these are probably stoliczkanus leyi moapae, nevadae, baileyi thermophilus) and Empetrichthys but DNA samples are only available from the Iraq popu- (latos concavus, latos latos, merriami, latos Pahrump, with all lations and not Iranian populations. A. ginaonis was only but latos latos being extinct now). The authors analyze the sampled from one location (Geno, Iran) and no surrounding relationships between those closely related taxa and assess stoliczkanus were sampled so it is unknown just how widely the validity of the subspecies designations given by Williams both species extend in Iran. and Wilde (1981). The new molecular results are It is clear, after reading this paper, that the taxonomy of the Aphanius species from Egypt to Iran is largely unsettled. We don’t know where Rüppell collected his samples of Aphanius dispar so extensive collection along the Red Sea and south- ern Israel are badly needed so as to redescribe of this species. More samples are needed from the area between Oman and Pakistan so as to better delineate the species. I expect many interesting research papers will grow out of Freyhof et al’s description of A. kruppi. [Tyrone Genade] Editor’s note: In this paper Freyhof et al. revalidate pre- vious synonym stoliczkanus with a new definition and raise richardsoni to full species status and it is acknowledged in Killi-Data, however more research is needed as to the final Photo of a pair of Crenichthys baileyi baileyi by Frans Vermeulen© identification of many populations of dispar s.l. in other re- gions and countries, including Iran, previously identified as notably solid because the 2 authors study 19 populations Aphanius dispar dispar. covering the entire distribution (and several specimens of [Jean Huber] each) and use the present state of the art of molecular tech- niques (nuclear sequences, mitochondrial sequences, haplo- Let’s jump in: A phylogenetic study of the great basin types and microsatellites markers) and they conclude that the springfishes and poolfishes, Crenichthys and Empetrichthys subspecies designations of Williams and Wilde are not valid (Cyprinodontiformes: Goodeidae). Campbell DC & Piller for Crenichthys, that rather a conservative approach suggests KR. PLOS ONE, 12:1–21, 2017. DOI https://doi.org/10.1371/ there are two species within nevadae and two species within journal.pone.0185425 baileyi but that no structure is found for latos. However the 2 This new comprehensive molecular study by 2 US re- authors do not go beyond that by naming the new molecular searchers from Southeastern Louisiana University focuses species based on the distinct population of nevadae (the north- on components of subfamily Empetrichthyinae (only 2 genera, ern clade) and by proposing a stable new systematic struc- with a relict distribution in southern central USA). This new ture for baileyi, probably because their results are somewhat study is very much welcome for the species belonging today to heterogeneous between mitochondrial and nuclear data and Crenichthys (baileyi albivallis, baileyi baileyi, baileyi grandis, bai- because they could not yet propose new diagnoses in line with

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 74 7 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 their results (etc.). In total, after a comparison of subresults The new N. cooperi refers to those populations from Mansa according to independently nuclear data, mitochondrial data (ZAM 97-2, ZAM97-3, ZAM 07-8, ZM 12-7) as well as Samfya. and haplotypes data, the 2 authors suggest that there are 2 to N. cooperi was fist discovered by John Rosenstock and Jorgen 4 distinct species in Crenichthys, and not 2 species, one with 5 Sunesen in 1989 in a small stream (the Lwingishi River, 36 km subspecies as previously, i.e., nevadae, the new un-named aff. east of Mansa). (On this same trip they discovered a female nevadae, baileyi and possibly moapae, while thermophilus and fish in the Luapula drainage near Kapalala which is today albivallis seem identical (and then each other synonyms and rosenstocki.) This species can be distinguished from sainthou- junior synonyms of baileyi). To go further (Kyle Piller pers. sei by a solid orange-red anal fin with irregular light blue zone comm. November 2017), the authors plan additional studies close to its base; and from rosenstocki by having a prepelvic (with more genetic results combined with new morphological length of 48.8–51.9% SL (vs 45.1–49%) and head depth of data) and those future studies are looked forward to, not only 75–77% HL (vs 78–84%) as well as a caudal peduncle length because of the conservation status of those species, but also 1.2–1.3 times it depth compared to 1.4–1.5 for rosenstocki and because the genetic distances between the discovered lineages sainthousei. The species could be discerned by a combination are low within Crenichthys based on cytb, however, this may of molecular phylogenetics, color pattern as well as principal be a trait common across the family Goodeidae and a result component analysis of meristic characters. of the recent estimated divergence time for Crenichthys. The habitat is described as a stream system composed of Additional note: a question related to this publication and multiple channels, flowing through dense grasses, roadside the subspecies concept has been raised to Killi-Data and an- ditches and residual pools. The water was always flowing swered in last November at: http://www.killi-data.org/faq. but the fish favored slow-flowing to still, densely vegetated php#Q(48): regions in the habitat. The pH was 6.18–6.35 and total dis- [Jean Huber] solved salts were less than 10–16 ppm and the water had a temperature of 24–26◦C. Other species of fish were present: Nothobranchius cooperi (Teleostei: Cyprinodontiformes): Enteromius barbs, Ctenopoma species as well as two Micropan- a new species of annual killifish from the Luapula River chax species. The conservation status of the species is given drainage, northern Zambia. Nagy B; Watters B; van der as Vulnerable. Merwe P; Cotterill F; & Bellstedt D. African Journal of Aquatic The species is named for Dr Barry Cooper who partic- Science, 42:201–218, 2017. DOI http://dx.doi.org/10.2989/ ipated in the collection of this species in 2007 (along with 16085914.2017.1372270. Brian Watters, Otto Schmidt, Bill Bishopp, Pieter Kearney, Nagy et al describe a new species of Nothobranchius. This Johan Jordaan and Johan Bornman), the specimens collected work is the product of a longterm study of Nothobranchius on that trip being pivotal to the phylogenetic analysis that species in Zambia, and follows on from the description of N. revealed this species. A report of the 2007 collecting trip is sainthousei from the same river system: the Luapula River given by Otto Schmidt (Auf Killi-Jagd nach Sambia. DKG- drainage. Keen readers of the sainthousei paper would have Journal 31 (4): 79-91, August 1999 and translated into English noticed that the rosenstocki populations were very genetically and published: 2008. Zambia revisited, 2007, Journal of the different. In this paper rosenstocki is split into rosenstocki and American Killifish Association 41:3–18). cooperi. [Tyrone Genade] https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 75 8 Genade: Killi-data News (Winter) Two new seasonal killifishes of the Austrolebias adloffi group from the Lagoa dos Patos basin, southern Brazil (Cyprinodontiformes: Aplocheilidae). Costa WJ; Cheffe MM; & Amorim PF. 2017. URL http://www.senckenberg.de/root/ index.php?page_id=16562#VZ67-2 This very detailed paper by Costa, Cheffe and Amorim describes 2 new Austrolebias species from the Lagoa dos Patos basin, in southern Brazil. The first is: Austrolebias pelotapes sp. nov. which is closely related to the other new species Austrolebias pongondo sp. nov. The clade comprising the two new species is supported as being more closely related to A. adloffi than to A. nigrofasciatus that is endemic to the same area. A. pelotapes n. sp. is distinguished from all other species of the A. adloffi group by having the urogenital papilla base attached by a thin membrane to the anterior margin of the anal fin in males. Austrolebias pelotapes seems to be the smallest species of the A. adloffi group, not reaching 35 mm SL (vs. maximum adult size between 43 and 45 mm SL). Austrolebias pelotapes is distinguished from A. pongondo by the former having 14–19 neuromasts in the supraorbital series (vs. 20–21) and 22 –23 neuromasts around orbit (vs. 24–27) and other features you may find in the description. Type locality: Brazil, Estado do Rio Grande do Sul: Município de Pelotas: temporary pool in the Sanga Funda drainage, tributary of Arroio Pelotas, 31◦43’46”S 52◦19’07”W; Collected by L. Lanés et al., 5 Nov. 2005. Austrolebias pongondo n. sp. is distinguished from all other species of the A. adloffi group, except A. pelotapes, by a combination of characters showing a transverse row of small spots on the middle portion of the dorsal fin in males (vs. transverse row absent in all other species), unpaired fins with a single row of light blue spots on their basal portion in males, From top to bottom: N. cooperi Mansa ZAM 97-3; N. rosenstocki Ka- and with a distinctive dark gray zone on the posterior portion palala ZM 12-9; N. sainthousei Mweshi ZM 12-19. Photos by Béla of the dorsal and anal fins and some other features mentioned Nagy©).

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 76 9 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 in the description. in the post otic series, shorter dorsal fin base length and in Type locality: Brazil, Estado do Rio Grande do Sul: Mu- females, having irregular spots arranged on the anterocentral nicípio de Pelotas: swamp at the road to Torotama island, Ar- portion of the flanks. raial, Povo Novo district, 31◦55’54”S 52◦14’42”W, Collected This species was found in the Pampa domain (a large by: M. Cheffe et al., 21 Aug. 2001. grassland) in pools that dry out in the dry season. It is One figure shows the Maximum likelihood tree of rela- found along both sides of the Rio Camaquã in small, shallow tionships among species of the Austrolebias adloffi group. Two pools located in grassland areas of farms. The plant life was tables show meristic data of both new taxa. One map shows mostly emergent and submerged plants. Other species of fish Geographical distribution of the group. Photos show life color were also captured: Austrolebias bagual, Callichthys callichthys, of A. pelotapes and preserved state. A. pogondo only in pre- Rhamdella eriarcha, Corydoras paleatus and Cheirodon ibicuien- served state. sis. The water is tinted brown but transparent, pH 5.71, salin- The paper is good reading and interesting is the discussion ity 0.04 ppt, conductivity 0.08 µS/cm, temperature 16.6◦C, in which, at the end, the need for clarifying the relationship dissolved solids 39 g/L and a depth of 25.2 cm. The sex ratio between A. salviai, A. reicherti, A. charrua and A. minuano of the species ranged from 1:1 to 1:2.4 males to females. The is recommended as a future study before synonymies being locations where the fish occur are already modified by man formally made. (rice fields are being cultivated, alien tree species have been [Frans Vermeulen ] planted and their is mining upstream). The authors recom- mend that this species be listed as critically endangered under A new annual fish of the genus Austrolebias (Cyprinodonti- caegory CR B2ab (ii, iii) of the IUCN. formes: Rivulidae) from Rio Camaq a basin, Laguna dos [Tyrone Genade] Patos system, Brazilian Pampa. Volcan MV; Gonçalves AC; & Lanés LEK. Zootaxa, 4338:141–152, 2017. DOI http: Description of two endangered new seasonal killifish species //dx.doi.org/10.11646/zootaxa.4338.1.7. of the genus Cynolebias from the São Francisco River The authors describe Austrolebias camaquensis. It was dis- basin, Brazilian Caatinga (Cyprinodontiformes, Aplocheil- covered in a temporary pool adjacent to the Arroio Santo idae). Costa WJEM. Zoosystematics and Evolution, 93:333–341, Antônio (a tributary of the Rio Camaquã). The pool is close 2017. DOI https://doi.org/10.3897/zse.93.20906. to highway BR-471, 30◦58’25”S52◦39’47”; and is within the In this paper, two new species of the genus Cynolebias are Canguçu muncipality, Rio Grande do Sul State, Brazil. It described from temporary pools of the Verde Grande River can be distinguished from other members of the Austrolebias drainage, São Francisco basin, northeastern Brazil. Described alexandri species group (except for paucisquama and juanlangi) are Cynolebias elegans sp. n., a member of the C. gilbertoi by having well-defined bright blue bars on its flanks in males. group, is considered as the smallest species of the genus, It differs from paucisquama by having contact organs on the reaching about 38 mm of standard length. The second is first rays of the pectoral fin and a greater number of scales Cynolebias gorutuba sp. n. belongs to the Cynolebias zeta-clade, along the caudal peduncle (14–16 vs 12). Its dorsal fin origin a group of large species supposedly feeding on smaller sym- is anterior to the vertical through the anal fin origin in males patric seasonal killifishes. The latter differs from other species compared to juanlangi as well as having fewer neuromasts of the group by the female color pattern, relative position of https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 77 10 Genade: Killi-data News (Winter) dorsal fin and vertebrae, and cephalic neuromast pattern. [Frans Vermeulen] Both species were not found in recent collecting trips, after their habitats had been drastically modified, and are possibly A new pelvic-less species of Melanorivulus Costa (Cyprino- highly endangered if not already extinct. dontiformes: Cynolebiidae), with a discussion on the pelvic- fin absence in killifishes. Deprá GDC; Silva HP; & Da Graça The author mentions two seasonal killifish generations WJ. Zootaxa, 4300:111–124, 2017. URL http://www.mapress. per year in most of the Caatinga region. The first one during com/j/zt/article/view/zootaxa.4300.1.6/0 the first rains (after the longest dry period), generally between In this paper the Brazilian authors describe a new species November and January, and the second one between February named Melanorivulus nelsoni which is found in a stream drain- and April. The author gives a clear overview of the history ing into the upper rio Paraguai basin, in the municipality of of the genus and its individual taxa. Amazingly all 16 now Juscimeira, State of Mato Grosso, Brazil. known species of Cynolebias from the Caatinga, except one, It is distinguished from all other congeners, except M. are described after 1990 due to recent in-depth sampling of pictus and M. planaltinus by the absence of pelvic fins and the temporary pools in that semi-arid area. The discussion girdle. The genus Melanorivulus overall does not show pelvic- focuses on the increasing rate of biodiversity loss and the high less taxa except for M. pictus and M. planaltinus which show risk of extinction of organisms in the Caatinga area mainly missing pelvic fin(s), one side or both, occasionally. Besides due to the high levels of specialization of these organisms. describing the new species in the present paper authors stud- The two new species provide a good example of two extreme ied the pelvic-less occurrence within the genus Melanorivulus. divergent morphological patterns within the genus Cynolebias. Authors present a clear history of the genus Melanorivulus, The small size and delicate morphology of C. elegans, with erected by W. Costa 2011 formerly in the genus Rivulus1, and relatively long fins in males, contrasts with the robust aspect describes the difficulties of determining the identity of sam- and short fins of C. gorutuba, which is a member of CZC ples if no life color information is available and when material (i.e. the Cynolebias zeta Clade), with species often reaching is available in preserved state only. over 100 mm SL or more and which predate on the smaller The two species M. pictus and M. planaltinus, both occa- Hypsolebias ssp they live with. sionally show missing pelvic fins, occur in nearby localities The name Cynolebias elegans refers to the Latin elegans (el- in the vicinity of Brasília, although in distinct river basins egant, fine), an allusion to the distinctive general appearance (respectively, upper rio Paraná and upper rio Tocantins). of the new species, with males combing relatively slender Melanorivulus nelsoni, in which all available specimens body with long unpaired fins. The name Cynolebias gorutuba lack both, the pelvic fins- and the girdle, was recently dis- refers to the Gorutuba River floodplains, the type locality of covered in the upper rio Paraguai basin, about 800 km from C. gorutuba. This paper is not only of great value for scien- Brasília and is described in the present paper. Additionally, tists and hobbyists but certainly for environmentalists and the authors discuss the possible genetic and evolutionary politicians who set the future for the region and must decide mechanisms behind the absence of pelvic fins in Melanorivu- how to protect the endangered ecosystem of the semi-arid lus and other Cyprinodontiform genera. M. nelsoni is named Caatinga. Very good reading! in honor of Joseph Nelson (1937–2011), who published on the

1KDI regards Melanorivulus as a subgenus of Rivulus.

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 78 11 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 absence of pelvic-fins in bony fishes earlier in 1989, and also Killifish Biology: Ecology & Physiology in recognition for his pivotal contributions to the knowledge of fish diversity as a whole. Effect of conservation of fish in formalin and ethanol on The paper is very informative and well written, with length-weight relationships and condition factor in Tlaloc clarifying tables(1), maps (1) and several photo clusters of labialis (Günther, 1866) specimen showing life colors, photo’s of museum material and . Anzueto-Calvo MJ; Velázquez- photo’s of the M. nelsoni habitat. Velazquez E; Matamoros WA; Cruz Maza BGA; & Nettel- A single point of criticism: The discussion around the Hernanz A. Journal of Applied Ichthyology, 33:1184—1186. occurrence of pelvic-less species in the genus Melanorivulus, DOI http://dx.doi.org/10.1111/jai.13461. like in the new species M. nelsoni but also occasionally in M. Fixation of fish with formalin and ethanol affects the vital pictus and M. planaltinus and other Cyprinodont fishes, is of statistics of the preserved fish. The formalin stiffens tissues great interest and, in my opinion, complex enough to bring and dulls colors while the ethanol, as well as washing out such in a separate paper and not include it in the description certain colors, also dehydrates the fish. This makes it difficult of this new taxon. Nevertheless the paper is excellent reading to use preserved fish for determining lengths, weights and for scientists and hobbyists. condition factors. In this paper Anzueto-Calvo et al set out to [Frans Vermeulen] determine how much the mass is lost due to preservation of Tlaloc labialis2.

Freshly collected fish were fixed in formalin for 72 hours and then moved to ethanol for various periods of time where after the weight and length of the fish was measured and compared to the initial measures of the freshly collected fish. Fomalin fixation caused a 5.78% loss of mass and 2.53% loss in standard length. A month in ethanol caused an 30% in mass. After 2 years the fish had lost 33.74% of the mass≈ and the fish’s standard length had reduced 3.47%. As consequence, freshly fixed fish give an over estimate of condition factors while long-term fixed fish will give an under estimate. This study provides a useful benchmark for future stud- ies of preserved specimens of T. labialis. These experiments would need to be repeated for each species to be able to use preserved specimens to determine condition factors. The author advised against comparing freshly collected and pre- served specimens. [Tyrone Genade]

2KDI lists this species as Profundulus labialis, Tlaloc is considered a subgenus of Profundulus. https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 79 12 Genade: Killi-data News (Winter) Do not eat your kids: embryonic kin recognition in an tus. Heffell Q; Turko AJ; & Wright PA. Journal of Compar- amphibious fish. Wells MW & Wright PA. Behavioral Ecol- ative Physiology B, Epub ahead of print, 2017. DOI https: ogy and Sociobiology, 71:140, 2017. ISSN 1432-0762, DOI //doi.org/10.1007/s00360-017-1123-4. 10.1007/s00265-017-2360-y, DOI https://doi.org/10.1007/ It is known that Kryptolebias marmoratus is able to respire s00265-017-2360-y through its skin. While emersed the fish are able to increase An intriguing title that proves to be more a statement than their body water content even though they are unable to drink a warning to future fish generations. The mangrove rivulus and are exposed to the air. The authors sought to find out (Kryptolebias marmoratus) are the investigated parents, due what changes occur to the skin tissue over time as the fish is to their ability to self-fertilize and produce isogenic strains emersed, and how this explains it ability to both respire and that can be easily differentiated. Long-term captive popu- gain water volume when emersed. lations originating from the USA (Florida) and Belize were Fish were acclimated to 15% or hypersaline 45% water utilized. Individual adults were kept well fed or starved for and then acclimated to air exposure on filter paper dampened two weeks ( to see if hunger played any role) and were then with water of 0.3, 15, 45%. Fish spent most of their time with placed in a container. An embryo was subsequently added their ventral skin in contact with the moist substrate but there and observations made on the time taken before the individ- was no particular skin area that permitted more water to pass ual investigated, the number of actual investigations and the across it into the fish. Freshwater acclimated fish had higher number of nips taken. The embryos were either from the water influx across the skin but after 7 days of emersion this parent, from the same strain or from a different strain. effect vanished. Skin thickness varied both with salinity and What they found was that no fish cannibalized its own em- air exposure. The dorsal skin thickness was unaffected by bryos but some did cannibalize those from the same strain and salinity but got thicker in response to air exposute. The ven- an increased number cannibalized those from other strains. tral epidermis was thicker in response to increasing salinity Interestingly there were differences between the two popula- and didn’t change when exposed to the air but the dermis did tions and the authors speculate on possible causes, indicating get thinner in response to emersion. areas for further research. The authors conclude that K. mar- The authors conclude that the fish are able to manage moratus adults can identify their own kin and differentiate their water balance by changing their skin thickness. Emer- on that basis when ‘considering’ cannibalism. An interesting sion increased skin water flux and the authors hypothesize paper that provides far more insight than can be reproduced that the fish are able to take up water when emersed due to here. The same volume of the journal features an editorial: the thinner ventral dermis. Kin recognition and filial cannibalism in an amphibious fish [Tyrone Genade] Joachim G. Frommen; Behav Ecol Sociobiol (2017) 71:141 DOI 10.1007/s00265-017-2369-2 that highlights the value of the Skeletal stiffening in an amphibious fish out of water is work as a first step in understanding filial cannibalism. a response to increased body weight. Turko AJ; Kültz D; [Andy Patel] Fudge D; Croll RP; Smith FM; Stoyek MR; & Wright PA. Journal of Experimental Biology, 220:3621–3631, 2017. DOI Plasticity of skin water permeability and skin thickness in https://doi.org/10.1242/jeb.161638. the amphibious mangrove rivulus Kryptolebias marmora- A very interesting paper but first watch this: https://www.

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 80 13 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 youtube.com/watch?v=mKxRe0hAQmg! To move on land The lack of a response to microgravity showed that the fish fish had to stiffen their skeleton for two reasons: the first were responsive to the body weight on the gill arches and not is that they needed firm support to propel their bodies and a side-effect of reduced buoyancy by emersion. The changes the second is that out of water they lose the buoyant sup- in collagen structure with little change in calcium deposition porting environment the water offerend and need to evolve a indicates that the fish stiffen their gill arches by changes in stiffer skeletal structure to maintain their body shape. A big the interactions between the collagen protein fibers. While question is whether weight-responsive bone is a fundamen- we may think that our bones are strong because they are hard, tal characteristic of bone or an early vertebrate evolved trait i.e. made up of calcium phosphate crystals, they are actually during the invasion of land. The goal of Turko et al’s study strong because of the collagen matrix in the bone. Without was to determine whether there is a cellular response by the this matrix the bone will crumble under pressure. That this skeleton of of amphibious fish in response to mechanical load fish, from an order of chiefly fully-aquatic species, can adapt when out of water or when the effect of gravity was artificially its skeleton to increased load when in a terrestrial state is reduced. interpreted, very cautiously, by the authors as evidence for a Two experiments were performed. (1) Fish were accli- weight-responsive bone being a fundamental characteristic of mated for 7 to 14 days out of water, and then half were reac- vertebrate bone. climated to water, and then the tensile strength of their gill [Tyrone Genade] arches (a skeletal tissue) was examined. (2) Fish were kept in an apparatus designed to reduce the gravitational force Environmental calcium regulates gill remodeling in a eury- on them (in this case to 0.06 of what we experience). Both haline teleost fish. Platek A; Turko AJ; Donini A; Kelly S; & water and terrestrial animals were exposed to microgravity. Wright PA. Journal of Experimental Zoology Part A: Ecological They made use of Picrosirius Red staining and Transmission and Integrative Physiology, 327:139–142, 2017. ISSN 2471- Electron Microscopy to assess changes in bone density; as well 5646, DOI http://dx.doi.org/10.1002/jez.2079. as proteomics to assess changes in gene expression in the bone This is an interesting paper with interesting implications of the gill arches. for the aquarium hobby. The gill is the site of more than just The gill arches of terrestrial-acclimated fish resisted more gaseous exchange, it plays an important role in nitrogenous force than gill arches of fish kept in water. This resistance waste excretion, acid-base regulation, osmoregulation and increased with time out of water, and decreased to normal ion regulation. For gaseous exchange the gill tissues need to levels when reacclimated to water. There were changes in the be thin, but for regulation of ions it needs to be thick. For abundance of various proteins in response to terrestrial accli- example, it is reported that goldfish under normal oxygen or mation, in particular to the levels of collagen and laminin and cool conditions will reduce their gill surface area to advantage other bone-related proteins. Histological analysis revealed an ion regulation; while under low oxygen or warmer conditions increase in collagen deposition and structure but no changes they will increase their gill surface area to increase gaseous were readily visible in the amount of calcium deposition in exchange at the expense of ion regulation. It is also known the bone. Fish exposed to reduce gravitation force didn’t un- that calcium ion concentrations affect gill surface area, in dergo any detectable modification to their skeletal structure particular the thickness of the layer of cells between the gill or skeletal strength. epithelium (exposed to the environment) and the endothe- https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 81 14 Genade: Killi-data News (Winter) lium (layer of cells that make up the blood vessels). This an interesting tie-in with Nothobranchius research. Does the layer is called the interlamellar cell mass and is observed in increasing dissolved solids of the pond signal the fish that the the channel between the gill lamellae. Platek et al hypoth- end is near and they have to increase reproductive rate at the esized that the gill surface area of K. marmoratus would be expense of their other tissues? modulated by water hardness. [Tyrone Genade]

K. marmoratus were kept in control water (hard well wa- Population dynamics of Melanorivulus rossoi, a restricted ter) or acclimated to low-calcium water (reconstituted RO geographic distribution killifish species. Severo-Neto F & Vol- water) or high calcium water (reconstituted RO water with can MV. Environmental Biology of Fishes, Epub ahead of print, extra calcium chloride added). After 7 days of acclimation 2017. DOI https://doi.org/10.1007/s10641-017-0695-x. fish were killed and fixed and their gills examined histolog- The authors set out to characterize the distribution and ically. As expected, the fish in the soft water had increased temporal variation in body size, sex ratio, density, length- interlamelar cell mass and therefore decreased gill surface weight relationship and condition factor of Melanorivulus area. rossoi3 in its natural habitat. As their motivation, the au- It is discussed in the paper that the move to soft water thors note that many of the recently described Melanorivulus could be stressful to the fish and the effect could be mediated have restricted distributions and very little is known about by increased cortisol. Even we, when stressed, will respond them other than they are vulnerable to extinction due to the by retaining cation such as calcium and sodium. As these small ranges. In this case, M. rossoi is threatened by landscape affects are also brought about by changes in temperature and modification for agriculture. oxygen availability it would be interesting to find out how Google Earth satellite images were used to look for suit- our cool stream inhabiting Aphyosemion respond to to oxygen able habitat for this species. The potential sites were surveyed levels and temperature. One of the symptoms of heat stress and, finally, a convenient site was chosen for monthly moni- in these fish is wasting, which would be expected from high toring. Specimens were anesthetized with eugenol (0.1 mg/L) cortisol levels and/or low oxygen levels. If our Aphyosemion before their vital statistics were taken. Specimens were re- respond to warm water by increasing gill surface area then turned to the wild once they had recovered from the eugenol. they would struggle to maintain their internal cation levels, Suspected predators were sampled (i.e. Hoplias malabaricus, causing them additional stress. This is an interesting avenue Synbranchus marmoratus and Gymnotus species) and their di- for research. Also, how the kidneys respond to the move from gestive systems inspected for M. rossoi. hard to soft water would also be interesting to know. In fresh- The paper contains a wealth of information and is inter- water fish the principle task of the kidneys is blood filtration, esting reading. salt retention and water excretion. The authors note that K. [Tyrone Genade] marmoratus often inhabit ephemeral pools in the mangrove forests and that the fish have to rapidly cope with changing Annual killifishes as model systems for advancing under- water conditions. Fish from more stable environments might standing of evolution and developmental biology. Podrabsky not be able to adapt as fast as K. marmoratus. This is also JE & Arezo M. Developmental Dynamics, 246:778–778, 2017. 3KDI regards Melanorivulus as a subgenus of Rivulus.

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 82 15 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 ISSN 1097-0177, DOI http://dx.doi.org/10.1002/dvdy.24594 manuscript provides a number of beautiful, clear photos that This paper is the editorial to the Developmental Dynamics will be useful in describing the morphological diversity of special issue “Annual Killifishes as Model Systems for Advanc- killifishes for years to come. ing Understanding of Evolution and Developmental Biology”. [Andrew Thompson] It is noteworthy for spelling out the need for an understand- ing of ecological and evolutionary consequences of altered Gene expression during delayed hatching in fish-out-of- developments; and a growing need to expand development water. Thompson AW; Hayes A; Podrabsky JE; & Ortí G. studies in the lab to more species. The authors note the multi- Ecological Genetics and Genomics, 3-5:52–59, 2017. DOI ple lineages present in the Cyprinodontiformes and the varied https://doi.org/10.1016/j.egg.2017.09.002. developmental trajectories. Fish-out-of-water provide unique opportunities for scien- [Tyrone Genade] tists to study the evolution and molecular mechanisms of stress response and adaptation to harsh environments. Study of the chorion of seasonal and non-seasonal Africa This study, which is rather technical, provides insight and Neotropical oviparous Cyprinodontiforme fishes. Mes- into how delayed-hatching embryos of Aplocheilus lineatus saddeq N; Hergueux J; Weickert JL; & Romand R. Environ- react to aerial incubation and suggest that delayed hatching mental Biology of Fishes, Epub ahead of print, 2017. DOI is a phenomenon distinct from the diapause stages of related https://doi.org/10.1007/s10641-017-0698-7. annual species. Similar patterns of gene expression are shared The study by Messaddeq et al. compares egg structure among Aplocheilus and other egg stranding and amphibious between the eight species and the laboratory model, Danio fishes. rerio (zebrafish). Eggs were imaged using scanning and trans- The methods of retrieving data are explained in detail mission electron microscopy. Both seasonal (Nothobranchius and a table with longest recorded pre-hatching delays in 43 patrizii, Nothobranchius guentheri, Spectrolebias filamentosus4, species from 8 different families is provided. Color figures Notholebias vermiculatus, and Austrolebias luteoflammulatus) show results of experiments but the reader is referred to the and non-seasonal (Fundulopanchax gardneri, Aphyosemion web version of this article for better view. striatum, and Anablepsoides5 rubrolineatus) species were exam- I think this paper is for specialists only, specialists that ined. Overall, the authors find that the structure of the egg like to understand more about the processes of delayed hatch- envelope is highly variable between species, but much more ing, diapauses and stress related environments that play a complex and ornate in killifishes than zebrafish. In addition, role in the survival techniques of our annual- and non-annual the egg envelopes are made of multiple layers that vary in Killi fishes. electron density, and different species possess envelopes with [Frans Vermeulen] differing thickness and numbers of layers. The authors report that egg structure is not necessarily correlated with ecology Females of the annual killifish Austrolebias reicherti (Cyp- or evolutionary relationships of the killifish species and state rinodontiformes: Rivulidae) recognize conspecific mates that further studies are needed to find such correlates. The based upon chemical cues. Blengini FR; Tassino B; & Pas-

4KDI considers Spectrolebias to be a subgenus of Simpsonichthys. 5KDI regards Anablepsoides as a subgenus of Rivulus. https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 83 16 Genade: Killi-data News (Winter) sos C. Behavioural Processes, Epub ahead of print, 2017. DOI Yamamoto’s solution and coco husk gave the best results https://doi.org/10.1016/j.beproc.2017.08.007. at 8 days (98.7% survival). Adding antibiotics gave a poorer In this paper Blengini et al report on experiments to deter- a result (91.7%). Distilled water and water with coco husk mine whether Austrolebias reicherti females recognize mates gave the worst results (72% survival). Eggs on Yamamoto’s based on chemical cues. The authors setup an aquarium that solution dampened cotton wool developed faster than those was subdivided into three sections. In this aquarium female in Yamamoto’s solution dampened coco husk. All eggs that reicherti were exposed to male A. charrua and reicherti. These exitted diapause developed uniformly in liquid medium. 50% two species are found sympatrically in the Río Cebollatí sys- of eggs placed in coco-husk water did not exist diapause II. tem but for the most past have a parapatric distribution. The Eggs kept in liquid medium had more eggs in diapause II authors selected fish from parapatric locations. They could at 100 days from laying than those which were moved onto either detect the fish of the opposite sex by sight or by smell. damp medium. Almost 80% of the eggs placed in coco husk Without exposure to the smell of the males the female re- water were still in diapause II after 100 days. icherti would associate with the male reicherti just as often as The authors speculate that something in the coco husk in- the male charrua, while if the female reicherti could use her terferes with embryogenesis. They interpret that the poor sense of smell she would select the correct mate, providing survival of eggs in distilled water was due to the proliferation strong evidence that these two species can distinguish each of fungi in the water. The authors conclude that Yamamoto’s other by scent. Data on female charrua mate choice wasn’t solution is an ideal medium for egg incubation of A. nigrofas- reported. ciatus. The solution is as follows: 0.75% NaCl, 0.02% KCl and [Tyrone Genade] 0.02% CaCl2. It is interesting that da Fonseca et al are able to induce diapause with coco-husk as effectively as Arezo (next Incubation media affect the survival, pathway and time article, below) was by keeping the eggs in peat in the presence of embryo development in Neotropical annual fish Aus- of the adults. In my experience with N. furzeri, eggs spawned trolebias nigrofasciatus (Rivulidae). da Fonseca AP; Volcan in sand and left with the parents will fully develop in three MV; & Robaldo RB. Journal of Fish Biology, Epub ahead of weeks. Could it be the peat that is inducing diapause and not print. DOI http://dx.doi.org/10.1111/jfb.13504. the parents? This is a very useful paper to hobbyists (as well as scien- [Tyrone Genade] tists working with annual killifish). It compares the effects of different incubation mediums on eggs survival and develop- Annual killifish adaptations to ephemeral environments: Di- ment. apause I in two Austrolebias species. Arezo MJ; Papa NG; The authors used distilled water (with or without pow- Berois N; Clivio G; Montagne J; & De la Piedra S. Developmen- dered coconut husk, i.e. coco peat), Yamamoto’s solution tal Dynamics, 246:848–857, 2017. DOI http://dx.doi.org/10. and Yamamoto’s solution with antibiotics (penicillin & strep- 1002/dvdy.24580. tomycin). For substrates the authors used cotton wool or Arezo et al provide an information-rich review of our cur- coconut husk or no substrate at all. The eggs were stored in rent knowledge of the regulation of Diapause but their main hermetically sealed containers at 20◦C in the dark. Survival focus was on characterizing diapause I at morphological and and development was monitored at regular intervals. molecular levels. As a prelude to this they began by evaluat-

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 84 17 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 ing the induction of Diapause I under laboratory and natural eggs must not be left with the adults for any length of time. conditions. To do so they set out to establish morphological On the other hand, if you want to maintain lots of different features of development of Austrolebias viarus and Austrolebias species but have few tanks, then by keeping the eggs with the charrua. adults the eggs can be forced into diapause I. The study by In brief, because the work was highly technical, eggs were da Fonseca (in this issue of KD-News) suggests that simply induced into diapause I through one of two incubation meth- moving the eggs from the peat water onto peat, exposed to ods. In one experiment freshly collected eggs were either the air, stimulates the eggs to develop to diapause II, and the incubated in Yamamoto’s Solution away from the parent fish, research on Millerichthys (in the previous issue) suggests that or sealed in a porous mesh inside glass bottles with peat moss emersion and redrying of diapause II eggs (and raising the sealed on the bottom with the parent fish. Oxygen levels were temperature) stimulates development to diapause III. 0.02 g/L in the bottles and all eggs were kept at 19◦C under a [Tyrone Genade] natural photoperiod. Eggs were checked for development on a regular basis. RNA was extracted at specific developmental Stem cells distribution, cellular proliferation and migration stages: epiboly, diapause IA (cells in dispersed configuration in the adult Austrolebias charrua brain. Torres-Pérez M; but rounded in shape), diapause IB (loosely arrayed rounded Rosillo JC; Berrosteguieta I; Olivera-Bravo S; Casanova G; cells in close proximity), reaggregation (exiting diapause I). García-Verdugo JM; & Fernández AS. Brain Research, 1673:11– 100% of eggs kept with the adults entered diapause while 22, 2017. DOI https://doi.org/10.1016/j.brainres.2017.08. 21% of the eggs kept in Yamamoto solution entered Diapause 003. I. A. viarius embryoes, compared to A. charrua, were more This paper continues previous work by the authors to prone to enter IA than IB. Embryos were monitored for 30 identify and characterize the neurogenic niches in the brain of days. The authors observed that the highest mortality occured Austrolebias charrua. In this paper they use a double-labeling in the blastula and gastrula stage. Viability was between 59% method to label fast and slow-cycling neurogenic cells to dif- in Yamamoto’s solution and 45% for embryos kept with the ferentiate the stem cells from the progenitor cells. adults. The alternative diapause state, IB, is reported by Arezo Using a combination of CldU and IdU they identified et al for the first time in this paper (but they note that it may fast cycling cells, slow cycling cells (i.e. the stem cells) and have been noticed by Levels et al in N. korthausae embryos) migrating cells in the brain of A. charrua. The migrating cells and they speculate that this could create more developmen- were observed moving out radially or tangentially from the tal diversity, and thus responses to environmental cues. It is stem cell niches. 10% of the labeled cells in all regions of the also observed that A. charrua exhibits high genetic variation brain were identified as stem cells. They used a 3D modeling compared to A. viarius. RNA quantities varied with develop- technology to digitally reconstruct the brains of the fish with mental state, first declining as the embryos enter diapause, locations of the labeled cells. and then increasing above the levels at epiboly as the embryo [Tyrone Genade] reaggregates. This paper is very useful to hobbyists eager to control Influence of temperature on age-related lipid peroxida- embryonic development of Austrolebias embryos. If the goal tion in a short-lived vertebrate (Nothobranchius furzeri). is rapid development and a quick second generation then Milinkovitch T; Lefrançois C; Durollet M; & Thomas-Guyon https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 85 18 Genade: Killi-data News (Winter) H. Fish Physiology and Biochemistry, Epub ahead of print, 2017. Nothobranchius guentheri. Lee Y; Drake AC; Thomas NO; DOI https://doi.org/10.1007/s10695-017-0439-z. Ferguson LG; Chappell PE; & Shay KP. Comparative Biochem- Previous research had shown an age-related accumulation istry and Physiology Part C: Toxicology & Pharmacology, Epub of lipid oxidation products (i.e. lipofuscin) in N. furzeri. In ahead of print, 2017. DOI https://doi.org/10.1016/j.cbpc. this paper Milinkovitch et al investigate the effect of temper- 2017.10.006. ature on the development of lipid oxidation products (lipid In this paper Lee et al report on the effect of resveratrol peroxides). supplementation on female fecundity. The authors fed fish The authors used the MZM 04-10 strain from the Chefu resveratrol and assayed egg production and egg fertility at 14 Rivier system and maintained them at either 22.1 or 25.6◦C and 20 weeks (as well as keeping track of the total number of and sacrificed fish at 7, 18 and 30 weeks of age. They then set eggs laid over 30 weeks). The fish fed resveratrol laid more about measuring the amounts of malondialdehyde content eggs over their lifespan and had higher egg fertility. At 20 of liver and muscle tissue. With age there was an increase weeks the resveratrol fed fish had larger ovaries and more in malondialdehyde in muscle tissue, with a sudden increase mature oocytes. Protein analysis of the ovaries for the rel- at 30 weeks. There was no difference with temperature in ative abundance of NAMPT and SirT1 revealed an increase muscle tissue samples. In the liver malondialdehyde levels in NAMPT in the resveratrol fish but not difference in SirT1 dropped from 7 to 18 weeks of age before increasing again at abundance. 30 weeks. The fish kept at 22.1◦C had more malondialdehyde NAMPT, Nicotinamide phosphoribosyltransferase, is an + in their liver at 30 weeks compared fish kept at 25.5◦C. enzyme involved in the regeneration of NAD which is needed The authors observe that their findings “disagree with the to maintain metabolism and Sirtuin activity, and accordingly, idea that oxidative stress varies positively with temperature.” the increase NAMPT activity could be extrapolated to an in- Liu et al showed the Austrolebias bellottii have a much faster crease in Sirtuin activity. The lack of SirT1 increase is at odds metabolism at cooler temperatures (Growth, 1975 39:337– with the work of Liu et al (Oncotarget. 2017, 8:55422–55434) 343) so an increase in liver metabolism and free radicals at which found an increase in SirT1 expression in the liver in re- lower temperatures isn’t unexpected but this is still a surprise sponse to resveratrol. It is already reported that the different result. Terzibasi et al showed an age-related increase of lipo- tissue of N. guentheri age differently (Dong et al, Fish Phys- fuscin in the from 9 week old liver of this strain (PloS One, iol Biochem 43:309, 2017, DOI 10.1007/s10695-016-0288-1). 2008 3:e3866) used by Milinkovitch so we should expect to Now it is reported that different tissues respond to resvera- see an increase in lipid peroxidation with age but this was not trol in different ways and this discovery needs to be taken observed in the liver and muscle. The authors speculate that into account when designing experiments to elucidate how the lower temperatures cause a decrease in oxidative stress resveratrol affects lifespan. in the lysosomes which generate lipofuscin. The disparity in [Tyrone Genade] results serves as an interesting departure point for further experiments into the physiology of Nothobranchius. The regeneration capacity of caudal fin in the common [Tyrone Genade] tooth-carp, Aphanius dispar (Rüppell, 1829)(Teleostei: Cyprinodontidae). Zeinali F & Motamedi M. International Dietary resveratrol increases mid-life fecundity of female Journal of Aquatic Biology, 5:321–327, 2017. URL http://

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 86 19 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 ij-aquaticbiology.com/index.php/ijab/article/view/375/0 thors recommend that glyphosate be regulated near aquatic This is the first investigation of fin regeneration in Apha- environment. nius. The authors used what they call Aphanius dispar from [Tyrone Genade] Bandar Abbas city in southern Iran. They kept the fish at Variations in fish body and scale shape among Aphanius three different temperatures: 23, 25 and 28◦C and at a pH of 6.8 to 7. The fish were fed a freeze dried live food and dry fish dispar (Cyprinodontidae) populations: insights from a geo- food. Fish were anesthetized with clove oil (180 ppm) for 3 metric morphometric analysis. Khosravi A; Golmakan MS; & minutes and then the caudal fin was cut down to the caudal Teimori A. Capsian Journal of Environmental Sciences, 15:113– peduncle. Fin regeneration was observed daily. Fin regenera- 123, 2017. URL http://cjes.guilan.ac.ir/article_2368.html tion was fastest at the higher temperature in both males and This is an interesting anatomical study of Aphanius dispar. females. The authors note that due to the short lifespan of the The authors were interested in the environmental and genetic fish this species could be used to study regeneration in aging. influence on fish morphology. They chose to study Aphanius [Tyrone Genade] dispar because it inhabits a broad range of habitats, shows a high tolerance to ecological and environmental changes and Evaluation of glyphosate toxicity on Arabian killifish, Apha- morphological variation from location to location. Also, many nius dispar collected from southwestern Saudi Arabia. Mes- populations are geographically isolated and some are translo- saad IA & Al Zailaie KA. Global Journal of Science: Frontier cations from small founder populations implying high genetic Research, 1:43–49, 2017. URL https://globaljournals.org/ divergence between populations. GJSFR_Volume17/4-Evaluation-of-Glyphosate-Toxicity.pdf. The authors collected fish from a man-made canal within Glyphosate, more popularly known as RoundUp, is in the Bandarabbas City, the Shur River (a brackish environment) news for its possible link to cancer. Messaad & Al Zailaie ex- and a man-made pool in Kahnuj city. Scale morphology was posed Aphanius dispar to varying concentrations of glyphosate studied using seven landmarks and a multivariate analysis and studied the consequences. to visualize shape changes mathematically was performed The authors experimented with several concentration whereby they could compare the shapes between populations. ranges: 0, 60–89, 90–119, 120–149 , 150–179, 180–209 and Morphological differences were observed between the pop- 210–240 mg/L. At the low dose the fish stopped schooling ulations and between the sexes and variance in morphology and became hyperactive with 20% mortality after 96 hours could be accounted for by the comparison of specific land- of exposure. All fish died at 250 mg/L doses. 96 hour LC50 marks. This data could be used to predict genetic relatedness. was calculated as 115.25 mg/L. Histological study revealed The authors note that changes in the fin and scale shapes abnormalities in the liver and gills of fish exposed to a quarter would effect the hydrodynamic power and swimming ability of the LC50 dose. of the fish. The authors conclude that the variation in body shape and scales among the populations are probably the For comparison, nile tilapia (Oreochromis niloticus) have an consequence of ecological conditions, in particular, to water LC of 1.05 mg/L and Tilapia zilli is 211.80 mg/L. In the 50 flow. USA glyphosate as been detected at 0.27–1.5 mg/L (http: [Tyrone Genade] //cdn.intechopen.com/pdfs/12592.pdf). There is little data on the long-term effects of glyphosate on animals. The au- https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 87 20 Genade: Killi-data News (Winter) The role of habitat choice in micro-evolutionary dynam- Fish were collected from the Maharlu Lake Basin, south- ics: An experimental study on the Mediterranean killifish western Iran and bred at Shiraz University. The fish were fed Aphanius fasciatus (Cyprinodontidae). Angeletti D; Seb- Artemia and Daphnia as well as artificial dry food. Methylene bio C; Carlini A; Strinati C; Nascetti G; Carere C; & Cim- blue was added to the water (0.4 g/L) of the aquaria to de- maruta R. Ecology and Evolution, Epub ahead of print. DOI ter fungus. Fish were bred in groups of 1 male to 3 females http://dx.doi.org/10.1002/ece3.3540. in tanks at 24◦C using green spawning mops. The authors The authors set out to sample A. fasciatus from different describe unfertilized eggs, fertilized eggs (1.4 mm in diame- microhabitats within the same location to determine if there ter), blastodisc formation, cleavage through the 2-cell stage to is any genetic divergence within the populations. the blastula formation, gastrulation, embryogenesis and then Fish were sampled in normoxic and hypoxic environments through to hatching. The entire process took 162 hours ( 7 ≈ and then compared the behavior between the fish under nor- days). The authors note that there was no sign of diapause in moxic and hypoxic conditions. The authors then analyzed six the development of the embryo. The photo micrographs are allozyme loci that were found to be highly divergent between beautiful. populations in previous studies. They found a connection [Tyrone Genade] between the glucose-6-phosphate isomerase-1 locus and the fish’s preference for and behavior under hypoxic or normoxic Phylogeography and population genetic analyses in the conditions. The authors state that this is evidence that indi- Iberian toothcarp (Aphanius iberus Valenciennes, 1846) at vidual behavioral traits and underlying genetics could lead different time scales. Gonzalez EG; Cunha C; Ghanavi HR; to nonrandom distribution of genotypes within a population, Oliva-Paterna FJ; Torralva M; & Doadrio I. Journal of Heredity, with some genotypes favoring certain microhabitats. Epub ahead of print, 2017. DOI http://dx.doi.org/10.1093/ Accordingly, habitat choice could then be a significant jhered/esx076. driver of micro-evolutionary dynamics and even speciation. Secondary freshwater fish species inhabiting fluctuating This observation could also affect molecular phylogenetics and extreme environments are susceptible to changes in dis- were few studies have been performed to determine whether persion, effective population size, and genetic structure. The the genes for sequencing have any effect on behavior or physi- Iberian toothcarp Aphanius iberus is an endemic cyprinodon- ological selection in the environment. tid of the Iberian Peninsula restricted to brackish water of salt [Tyrone Genade] marshes and coastal lagoons on the eastern Spanish Mediter- ranean coast. The genetic structure, diversity and historical Towards the conservation of a critically endangered species, demography was studied analyzing mitochondrial and nu- Aphanius farsicus: embryogenesis and development. Va- clear markers across its distribution range. The results indi- hed NS; Esmaeili HR; Masoudi M; & Ebrahimi M. Environ- cated a subtle levels of phylogeographic structuring. This, mental Biology of Fishes, Epub ahead of print, 2017. DOI combined with substantial mitochondrial diversity, suggest https://doi.org/10.1007/s10641-017-0691-1. that Pleistocene glaciations had a lesser effect on the demo- The authors describe the embryonic development of Apha- graphic structure of its populations than was the case for nius farsicus. This is the one of the first descriptions of em- Iberian fresh-water species with similar distribution. Other bryogenesis of Aphanius species. test showed a relatively high degree of mitochondrial differ-

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 88 21 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 entiation, which could be explained by historical isolation of a population. Conversely, significant genetic differentiation, following an isolation-by-distance pattern was detected with microsatellites, suggesting extensive habitat fragmentation over the past hundreds of years. At a smaller geographical scale, habitat fragmentation, probably due to human activity, appears to have resulted in substantially reduced migration and increased genetic drift, as shown by expanded genetic differentiation of populations. [Stefano Valdesalici]

Shoal sex composition affects exploration in the Mediter- ranean killifish. Lucon-Xiccato T & Griggio M. Ethology, Epub Photo of a male Aphanius fasciatus from Capu Biancu FRN 10-12. ahead of print. DOI http://dx.doi.org/10.1111/eth.12654. Photo by Béla Nagy©. A paper by the same authors was reviewed in the last issue of KDN and appears based on the same experimental meth- Aphanius farsicus Teimori, Esmaeili & Reichenbacher, 2011 ods involving Aphanius fasciatus. In this paper the authors is an endemic fish restricted to the Maharlu Lake basin in Fars go further in analysing the results and highlight that females Province, southwestern Iran. Keivany and Esmaeili suggested seem to have a far greater effect on the behavior of a mixed that the species should be in the IUCN’s Red Data Book due to sex shoal than males. In making comparison to studies on criteria such as restricted distribution, destruction of spawn- other taxa the authors discuss different selective pressures on ing grounds, environmental pollution and drought. There the sexes, the effect of individuals on group behavior and in are several scattered patches of this fish species in spring particular how this might impact on exploration. They urge systems around the Maharlu Lake basin isolated from each more use of groups of fish to investigate exploratory behavior other by land or by a hyper saline lake. The authors inves- pointing out that many studies have previously focused on tigated genetic structure from four different water bodies in individuals only. A practice that may well prove less than the Maharlu Lake basin applying five microsatellite markers. satisfactory. The results are a high genetic diversity observed within the [Andy Patel] populations (99%) and low diversity (1%) among them indi- cating probably high level of gene flow among the studied Assessment of genetic diversity of an endangered tooth- populations at the present time or in the past. The reasons carp, Aphanius farsicus (Teleostei: Cyprinodontiformes: can be hypotheses of annual floods around the lake make a Cyprinodontidae) using microsatellite markers. Yaripour fresh water area in some parts of it, and fishes can be migrated S; Esmaeili HR; Gholamhosseini A; Rezaei M; & Sadeghi S. from one spring to another. Secondly the currently isolated Molecular Biology Research Communications, Epub ahead of populations may be relicts of a recently widespread species print, 2017. DOI https://doi.org/10.22099/mbrc.2017.24404. around the Maharlu Lake and therefore, isolated populations 1246. did not have enough time for differentiation and form part of https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 89 22 Genade: Killi-data News (Winter) a single genetic population. might be a molecular target for thermal adaptation in other [Stefano Valdesalici] species. [Tyrone Genade] Intraspecific variation and plasticity in mitochondrial oxy- gen binding affinity as a response to environmental tem- Preliminary Insight into Winter Native Fish Assemblages perature. Chung DJ; Morrison P; Bryant H; Jung E; Brauner in Guadiana Estuary Salt Marshes Coping with Environ- C; & Schulte P. Scientific Reports, 7:16238, 2017. DOI https: mental Variability and Non-Indigenous Fish Introduction. //doi.org/10.1038/s41598-017-16598-6 Gonçalves R; Cruz J; Ben-Hamadou R; Teodósio MA; Correia Mitochondria supply the energy needs of cells. Most stud- AD; & Chícharo L. Fishes, 2:19, 2017. DOI http://dx.doi.org/ ies focus on the maximum capacity of mitochondria under 10.3390/fishes2040019. certain environmental conditions and extrapolate those data While the Mummichog may be facing some pressure in to hypoxic and thermal conditions. In this paper Chung et al Delaware Bay (see previous review) in the Guadiana estuary investigates mitochondrial oxygen kinetics at submaximum of South-East Portugal it is the Mummichog applying the pres- capacity and how it could contribute to local adaptations and sure. The authors surveyed an extensive fish fauna across the plasticity in response to changes in temperature. estuary and found that in certain parts environmental degra- The authors used two subspecies of Fundulus heteroclitus dation is leading to extreme conditions in certain areas that subspecies for these experiments. They acclimated the fish favour just two species, a native goby and the non-indigenous to 15◦C and studied how oxygen consumption rates at 50% Mummichog (between them comprising over 99% of catches). of the mitochondrial capacity. They found that the southern F. heteroclitus and many other species have been introduced by subspecies is more tolerant of thermal stress and hypoxia humans and though many have fared poorly the Mummichog, and had higher oxygen affinity compared to the northern sub- with its high adaptability and tolerance, appears to be on the species. Acclimation to temperature extremes (5 and 33◦C) edge of gaining a foothold. Various sizes of individual were altered the oxygen affinities of the mitochondria for both sub- found, from 1.6 mm up to 52 mm, suggesting local breed- species. At 33◦ the southern and northern subspecies didn’t ing. The estuarine fish fauna may therefore be at risk and the show any difference in mitochondrial oxygen affinity. The authors urge further monitoring of the area. northern fish had lower oxygen affinity in their mitochondria. [Andy Patel] In acute exposure to thermal stress the northern subspecies still had lower oxygen affinity. Blood oxygen affinity was also Ryanodine receptor and FK506 binding protein 1 in the At- investigated but, interestingly, showed no difference under lantic killifish (Fundulus heteroclitus): A phylogenetic and the temperate conditions. population-based comparison. Holland EB; Goldstone JV; Pes- The authors report that there were clear effects due to sah IN; Whitehead A; Reid NM; Karchner SI; Hahn ME; Nacci genetic background and environmental acclimation on mito- DE; Clark BW; & Stegeman JJ. Aquatic Toxicology, 192:105 – chondrial oxygen affinity. The higher oxygen affinity observed 115, 2017. DOI https://doi.org/10.1016/j.aquatox.2017.09. in the southern subspecies are probably an adaptation given 002. that they are from a warmer climate and more likely to ex- Mummichogs once again prove their worth as a model perience oxygen deprivation due to thermal stress and this species for research into the effects of pollution on physio-

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 90 23 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 logical pathways. This study looks at the effect of widely is not attributable to reduced prey, rather to unknown other encountered non-dioxin like polychlorinated biphenyls (NDL factors. This leads the authors to warn against traditional PCBs) and their neurotoxic effects. By studying two popu- armoring methods, which can adversely affect native fauna, lations of Mummichog, one tolerant and one intolerant the and urge use of alternative methods for shoreline protection. authors identify six genes that activate Ryanodine receptors [Andy Patel] (intracellular Ca2+ channels). They point to the relevance of their work in establishing a framework against which to Zinc chloride rapidly stimulates efflux transporters in re- compare other teleosts and mammals and their responses to nal proximal tubules of killifish (Fundulus heteroclitus). NDL PCBs. Perhaps of some interest to killifish researchers Zaremba A; Miller DS; & Fricker G. Toxicology and Applied but of greater interest to toxicolgists. Pharmacology, 334:88 – 99, 2017. DOI https://doi.org/10. [Andy Patel] 1016/j.taap.2017.09.001. The authors set out to characterize the effect of ZnCl2 on Growth and movements of Mummichogs (Fundulus hete- the activity of Multidrug resistance-related protein 2 (Mrp2). roclitus) along armored and vegetated estuarine shorelines. Zinc is an essential nutrient that is needed to fight infection, Crum KP; Balouskus RG; & Targett TE. Estuaries and Coasts, modulate inflammation, male fertility and maintain nervous Epub ahead of print, 2017. DOI https://doi.org/10.1007/ system function (cognition, sensory systems and mood). Mrp2 s12237-017-0299-x. is involved in the excretion of metabolic toxins and xenobi- A study to determine the impact of urbanization on the otics (eg. antibiotics) across the cell membranes of the renal widely distributed Mummichog in the Delaware Coastal Bay tubules cells into the urine but is also present on the cells of Area. The authors use tagging and recapture to measure lung, gut, liver and blood-brain barrier. Zaremba et al chose growth rates and movement of individuals along 4 differing to experiment on Fundulus heteroclitus as it is a well estab- types of shoreline, two armored by humans to stabilise the lished model organism for studying the effect of chemical shoreline and two naturally vegetated. The armored comprise toxicants and pollutants. bulkheads and riprap (stone aggregate) and the vegetated are Zaremba et al dissected renal tubule tissue from freshly marsh areas, populated by Spartina alterniflora and Phragmites killed fish and then exposed them to ZnCl2 or CdCl2 to de- australis. Mummichog show strong site fidelity in natural termine their effect on ion flux across the tubule cell mem- environments and this behavior is replicated in all 4 shoreline branes. The movement of ions accross the membrane was types with most specimens recaptured within 5 metres of measured using a florescent dye, Texas Red. ZnCl2 increased their original tagging site. Only a few specimens overcame flourescent labeling in the kidney tubule lumen about 5 times their natural stay-put tendency and traveled to different area while CdCl2 had only a small (but significant) effect. The types, mostly from armored to vegetated. Perhaps unsurpris- effect of the ZnCl2 increased with ZnCl2 dose. This effect ingly the armoring shows a negative impact on Mummichog was not affected by stopping protein synthesis with cyclohex- numbers when compared to marsh areas. However growth of imide but was stopped by inhibition of the endothelin recep- the reduced number of individuals increased in armored ar- tor type B/nitric oxide synthase/protein kinase C signaling eas, but not sufficiently to overcome the negative overall effect pathway. Inhibitors of phosphatidylinositol 3-kinase activity, on productivity. So it appears that the decrease in population glucocorticoid-inducible kinase 1 and mTOR (e.g. rapamycin) https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 91 24 Genade: Killi-data News (Winter) also abolished the effect of the ZnCl2. ZnCl2 also stimulated Roberts BJ. Environmental Biology of Fishes, 100:1315–1327, P-glycoprotein and Breast cancer resistance protein. 2017. DOI https://doi.org/10.1007/s10641-017-0645-7. From a biochemical perspective I find this a very interest- The authors investigated populations of ing paper. Why would ZnCl increase the activity of Mrp2 Fundulus gran- 2 in marsh areas of the northern Gulf of Mexico. Initially to pump Texas Red out of the cells into the lumen? Is there dis fish were captured and tagged. They were bathed in oxyte- an optimum level of Zn2+ needed to faciliate proper kidney tracycline to leave discernible marks on their otoliths and function, to pump out xenobiotics and other metabolic tox- released. When subsequently recaptured, either in April (to ins? In very soft aquarium water, would our fish benefit from determine winter growth rate) or September (to determine some zinc in the water? Are their diets sufficient in zinc for summer growth rate) the otoliths were removed and exam- fish health and fertility? Fish need 15–40 mg/kg feed (Watan- ined microscopically. Results supported the validity of using abe et al. 1997. Aquaculture 151:187–207) and most feeds otoliths to measure ageing, as winter growth produced a more have zinc in excess. For example, black soldier fly larvae opaque area than summer. This allowed them to identify in- have 108 mg zinc per kg (dry mass) and earthworms have 198 dividuals as under 1 year, more than 1 year and more than mg/kg while shrimp has only 15 mg/kg. High levels of copper, 2 years and then refine the age estimates by measuring the phosphorus, calcium and phytates inhibit zinc uptake by the actual amount of growth of the otoliths between first and gut (http://www.fao.org/docrep/field/003/ab470e/ab470e06. second capture. The oldest fish was estimated at 2.25 years ( htm) which could mean problems for our softwater fish kept 0.25). ± in hard water or water full of phytates. Experiments on rain- bow trout showed that dietary supplementation with zinc Growth of the otoliths was fastest in summer, as might be enriched yeast did improve immune function (Gharekhani et expected. Growth of the actual fish was also recorded (body al, 2015. Iran J. Vet. Res. 16:278–282). It might be possible length) and this actually showed significantly faster somatic to enrich Artemia but feeding baby grindal worms (which are growth in winter. This is in contrast to previous findings in Annelids like earthworms) might be an easier option. Sadly no F. grandis. The authors comment on the fact that studies of data is available on zinc content of Artemia or grindal worms reproductive activity in Gulf Killifish show reductions as tem- but there is evidence of the latter accumulating zinc in its tis- perature rises (stopping at 30◦C), which may correlate with sues from the environment—as much as 458 mg/kg in worms growth. Recorded temperature at the trial areas ranged be- from polluted flood plains in the Netherlands (Van Vliet et tween 1.7◦C and 34.7◦C with the summer mean close to that al. 2006. Euroean Journal of Soil Biology, 42:S117–S126) sug- 30◦ cutoff. As they point out this is in marked contrast to the gesting that grindal and white worms could be enriched by Atlantic Killifish (F. heteroclitus), a predominantly temperate feeding them foods rich in zinc or adding a small amount of species whose growth increases with temperature. Despite zinc chloride directly to their growth medium. their similarity F.heteroclitus also live longer (upto 4 years). [Tyrone Genade] With age and growth rates now discernible for F. grandis Age validation and seasonal growth patterns of a subtropi- the authors promote its potential as an indicator species in cal marsh fish: The Gulf Killifish, Fundulus grandis. Vastano the marsh areas of the Gulf. AR; Able KW; Jensen OP; López-Duarte PC; Martin CW; & [Andy Patel]

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 92 25 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 Salinity and temperature effects on element incorpora- cided to compare behaviours between the captive and ‘wild’ tion of Gulf Killifish Fundulus grandis otoliths. Nel- populations. As of 2013 each population had passed through son TR; DeVries DR; & Wright RA. Estuaries and Coasts, 15 generations and indeed there were clearly discernible dif- Epub ahead of print, 2017. DOI https://doi.org/10.1007/ ferences between them. They introduced captive fish to a s12237-017-0341-z. separate pool from the wild fish and made observations to The chemical analysis of otoliths for various assays rely on see if there were any actual behavioral differences that might the ratio of strontium to calcium and barium to calcium. The have implications for future repopulation efforts. To mitigate authors set out to determine how salinity and temperature against habitat differences they also dug out similar area in affect these two chemical ratios. each locale so that they could observe directly comparable for- They collected Fundulus grandis from various locations aging, mating and territorial behaviors albeit with a limited in the northern Gulf of Mexico. The ratio of strontium to number of fish. calcium increased with temperature and salinity. Conversely, They found that despite genetic divergence none of the the ratio of barium to calcium decreased with salinity but key behaviors were materially affected. They note that this increased with temperature. may be a positive sign for ongoing and future efforts to pre- The authors caution that temperature variability can con- serve extirpated species by captive maintenance. found extrapolation of past salinity from otolith analysis and [Andy Patel] recommend using both the strontium to calcium and barium to calcium ratios for reconstructing fish environmental his- tory. [Tyrone Genade]

Preservation of behavior after fifteen years of isolation: comparisons of wild and captive endangered pupfish in their natural habitat. Black A; Snekser JL; & Itzkowitz M. Envi- ronmental Biology of Fishes, Epub ahead of print, 2017. DOI https://doi.org/10.1007/s10641-017-0662-6. The Leon Springs pupfish, Cyprinodon bovinus having been extirpated in the wild has been subject to conservation mea- sures involving maintaining a relatively large captive popula- tion. Between 1998 and 2001 captive fish were reintroduced in an effort to repopulate its native environment. Most quickly died out but a small number (just 1 breeding male amongst them) were assisted by habitat regeneration and persist as a small population. Noting that captive populations can un- dergo genetic and morphological changes that can render them incapable of adapting to reintroduction the authors de- https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 93 26 Genade: Killi-data News (Winter) Posters & Conference Abstracts Theses & Dissertations

Nothobranchius furzeri: A new model organism of alpha- Integrating Physiology and Genomics to Identify Mecha- synucleinopathy. Genade T. J Alzheimers Dis Parkinson- nisms of Adaptation in Killifish. Brennan RS. , University ism, 7:5 (Suppl), 2017. DOI https://doi.org/10.4172/ of California, Davis, 2017. URL https://search.proquest. 2161-0460-C1-029. Presentation at the 3rd International com/openview/5fdef32b308743b3efea0272551b7809/1? Conference on Parkinson’s disease and Movement Disorders, pq-origsite=gscholar&cbl=18750&diss=y Sept 25-26, 2017, Chicago, USA. Fundulus heteroclitus has diverged from marine to fresh wa- ter several times and has produced genetically distinct, locally Annual killifish, Nothobranchius furzeri, discloses Parkin- adapted populations. The authors set out to understand the son’s disease phenotypes. Matsui H. Journal of the Neuro- mechanisms enabling the evolution of this fish into diverse logical Sciences, 381:580 (Suppl), 2017. DOI https://doi.org/ habitats. The author integrated physiological, transcriptomic 10.1016/j.jns.2017.08.1634. Presented at the World Congress and genomic mechanisms to identify mechanisms contribut- of Neurology (WCN 2017), September 16–21, 2017, Kyoto, ing to evolutionary divergence. The author identified gene Japan. loci that are under divergent selection. In brief, both Prof Matsui and I have been exploring the [Tyrone Genade] feasibility of N. furzeri as a model of Parkinson’s Disease. In 2013 I discovered that the protein α-synuclein accumulates in Effects of embryonic arsenic exposure on killifish (Fundulus the brains of N. furzeri Gonarezhou with age. This is the patho- heteroclitus) growth, feeding behavior, and intestinal mor- logical protein in Parkinson’s and several other neurological phology and cell types. Sims KC. Masters thesis, Clemson Diseases. Prof Matsui has confirmed this observation as well University, 2017. URL https://tigerprints.clemson.edu/all_ as the loss of dopaminergic neurons in the motor system of N. theses/2741 furzeri (among several other discoveries). In my presentation A thesis that makes use of the popular model fish Fundulus I present histological evidence for α-synucleinopathies in the heteroclitus to examine whether arsenic exposure in human brain of the fish as well as evidence for the dysfunction of DJ- fetal development is likely to cause long term harm and deter- 1, the protein coded by the PARK7 gene which is implicated mine ways that might occur. Mummichog are chosen for their in human familial Parkinson’s Disease. It is a remarkable prolific egg production (increasing number of test results) coincidence that we both presented our work at almost the their similar reaction to arsenic exposure and of course their same time. We had no idea what each other were doing. much shorter life cycle. A clear and logical literature review [Tyrone Genade] is presented establishing the basis for the study and reasoning for the subsequent experimental investigations. Experimentation involved exposing embryos to different arsenic concentrations and then examining the persistence of three effects; alteration of feeding behaviour, disruption of nutrient uptake, reduction in insulin dependent growth factor (IGF). While all three are affected in the short term, it is

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 94 27 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 the latter that appears to have the most detrimental long-term able to show changes in polycyclic aromatic hydrocarbons effect in that it probably reduces condition and growth during metabolism and immunology within two generations. the (pivotal) juvenile stage. This is a very rapid adaption to environmental changes! Of some interest to killifish enthusiasts the thesis is pre- It seems that Fundulus heteroclitus isn’t the only member of dominantly focused on examining the potential dangers to hu- this genus that is able to rapidly evolve to adapt to cope with mans of low level arsenic exposure and concludes that current negative environmental changes. But perhaps this isn’t un- recommended levels may need to be lowered. Nevertheless expected. Molecular phylogenies place F. grandis as sister the subject is fascinating and for those with an interest in the species to F. heteroclitus. effects of low level exposure to toxic substances (regardless of [Tyrone Genade] animal group) a recommended read The annual killifish Nothobranchius furzeri as a new [Andy Patel] model in long-term ecotoxicological research. Philippe Ed’s note: Arsenic is a common contaminant of ground C. , Katholieke Universiteit Leuven, 2017. URL https:// water. The author reported affects from 10 µg/L. Aquarists lirias.kuleuven.be/handle/123456789/599381?mode=full& drawing water from wells for their fish should keep this in submit_simple=Show+full+item+record mind. See https://water.usgs.gov/nawqa/trace/arsenic/ for This thesis isn’t available for reading. The abstract indi- USA data on arsenic concentrations in ground water. Simi- cates that much of it is based on Philippe et al’s previous lar results exist for the UK: http://www.bgs.ac.uk/research/ paper on the use of Nothobranchius ecotoicology (Philippeet highlights/2013/arsenicSW.html. If you are using well water al, Ecotoxicology and Environmental Safety, 144:26–35, 2017. you should have it tested. DOI 10.1016/j.ecoenv.2017.05.047). The abstract indicates [Tyrone Genade] that additional experiments were done testing the interaction Multigeneration effects of chronic exposure to polycyclic between toxicant and temperature as well as the effect of sub- aromatic hydrocarbons in Gulf Killifish (Fundulus gran- lethal concentrations of chlorpyrifos. It would appear that dis). Yammine A. Masters, Louisiana State University at several interesting papers are still to come from this research. Baton Rouge, 2017. URL https://digitalcommons.lsu.edu/ [Tyrone Genade] gradschool_theses/4327/ The author set out to examine potential adaptation within two generation of gulf killifish exposure to polycyclic aro- matic hydrocarbons. They also tried to determine whether mu- cus could be used to determine levels of 11-ketotestosterone and vitellogenin levels in gulf killifish. Both mucus 11-ketotestosterone and vitellogenin levels correlated positively with blood levels of these two substances; and both were responsive to exposure to pollutants. Expo- sure to polycyclic aromatic hydrocarbons causes a drop in sperm motility and oocyte cell wall thickness. The author was https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 95 28 Genade: Killi-data News (Winter) Interesting research on other fish ogy, Epub ahead of print, 2017. ISSN 1439-5444, DOI https://doi.org/10.1007/s10164-017-0523-y An interesting paper, with video links embedded, looking Understanding the effects of temperature on sex ratio at the degree of mating behaviour vs aggressive behaviour in a sexually dimorphic fish species. Chance DL. Senior between males and females of a Costa Rican species. The thesis, Universty of California, Santa Cruz, 2017. URL authors provide commentary on the current state of research https://norriscenter.ucsc.edu/natural-history-resources/ into mating and aggressive behaviours in livebearers and iden- student-created-resources/index.html tify some previously undocumented behaviours in B. olom- Chance explores the connection between ambient temper- ina. They also note that behaviours can vary widely between ature and sex ratios. It is theorized that an increase in ambient species and draw attention to the need to study multiple temperature would result in more males to females because species in order to elucidate how such behaviours have devel- the increased temperature exacerbates the metabolic costs to oped. Their study does not include male-male interactions females, which need to invest more energy into the production but in male-female interactions they do see evidence that the of eggs, over males. males can detect female breeding condition and from some Chance surveyed 20 geothermal sites in California and distance. No female behaviour was noticed that might specifi- New Zealand. The author controlled for the fish’s temperature cally account for this though they note the short trial duration preference in open habitats where a temperature gradient was may not have allowed time for the females to demonstrate it. present. The study showed that across the sites studied the Well referenced and raising many questions on male-female ratio of males to females increased with an increase in tem- interactions that require further study. perature. Interestingly, within open systems the sex ratio is [Andy Patel] closer to 1:1 than in closed sites and more females were found in the warmer regions of the habitat. The author speculates The protected areas system in Brazil as a baseline condition that the gravid females favor the warmer sites in such open for wetlands management and fish conservancy: the exam- environments. The observation of spatial variation in sex ra- ple of the Pantanal National Park. Polaz CNM; Ferreira FC; tio has consequences for future studies of how temperature &Junior˜ MP. Neotropical Ichthyology, Epub ahead of print, effects sex ratios in the environment. 2017. DOI http://dx.doi.org/10.1590/1982-0224-20170041. No controlled studies were conducted in aquaria. It would This paper is a report on the Pantanal National Park and be interesting to know whether the effect is preserved under how it is being used as a baseline by which to measure biotic captive conditions, and whether all populations respond the integrity in other systems. The authors focused on the fish same way to increased or decreased temperature. fauna of the park. The authors studied four categories of [Tyrone Genade] aquatic habitat: channels in the flood plains, permanent bays, temporary bays and rivers. 146 fish species were identified Mating and aggressive behaviour of Brachyrhaphis olom- and fish sampling occurred during the dry season. There was ina (Cyprinodontiformes: Poeciliidae). Garita-Alvarado no relationship between the habitat category and fish diversity. CA; Naranjo-Elizondo B; & Barrantes G. Journal of Ethol- Only one killifish was identified: Melanorivulus punctatus6. 6KDI regards Melanorivulus as a subgenus of Rivulus.

PublishedKilli-Data by NWCommons, News, Volume 2017 2, number 4, Winter 2017 96 29 Killifish Research Review, Vol. 2 [2017], Iss. 4, Art. 1 [Tyrone Genade] as Fundulus diaphanus; the pre-optic area, the dorsal section of the ventral telencephalon, the ventral section of the same Are fancy guppies (Poecilia reticulata Peters, 1859) in- telencephalon, the supracommissural part of the ventral pal- vasive? Bandaranayake C & Chandrasekara W. Sri Lanka lium. Their findings confirmed previous studies in which Journal of Aquatic Sciences, Epub ahead of print, 2017. DOI guppies showed marked preferences for social interaction and http://doi.org/10.4038/sljas.v22i2.7536. a preference for larger groups. However the assay techniques The simple answer is no. Fancy guppies can’t compete, only revealed changes within the pre-optic area. They specu- at least not against the wild type forms they were compared late on potential reasons as to why this might be and argue for to in experiments. The paper doesn’t gage the ability of the more research into the brain biology of the important model fancy guppies to compete with native species. One thing not species—the guppy. considered is that fancy strains spontaneously regress to the [Andy Patel] wild type form. [Tyrone Genade]

Forebrain activation during social exposure in wild-type gup- pies. Cabrera-Álvarez MJ; Swaney WT; & Reader SM. Physiol- ogy & Behavior, 182:107–113, 2017. DOI https://doi.org/10. 1016/j.physbeh.2017.10.012. The authors undertake two experiments with wild type Trinidadian guppies to determine whether social interaction can be detected by changes in immediate early gene expres- sion within four areas of the brain. By exposing individual guppies to the stimulus of no (zero), small (two) and large (ten) shoals and also by studying preferences for individuals with no, small and large shoals. Using gene assay techniques they looked for activity within regions of the brain known (or analagous to those known) to contribute to the social decision making network in other vertebrates and fish species such Photo by Hristo Hristov©

Killi-Data News, ISSN 2495-330X. Periodicity: quarterly. Publication: Killi-Data Editions, Paris. Address : Killi-Data International, Jean H. Huber, 7 Boulevard Flandrin, 75116 Paris, France ([email protected]). Date: 23 December 2017. Price per issue: e 3 Citation : Killi-Data News, Winter 2017, 2 (4) (December 23): 30 pp. (2017-Killi-Data_News-December-vol2n4-ISSN2495-330X.pdf)

https://nwcommons.nwciowa.edu/killiresrev/vol2/iss4/1Killi-Data News, Volume 2, number 4, Winter 2017 97 30